Phenoxyalkylcarboxylic acid derivatives in the treatment of irritable bowel syndrome

ABSTRACT

A method of treating irritable bowel syndrome in a patient suffering therefrom comprising: administering an effective amount of a compound selected from compound 1, its metabolite 2 and pharmaceutically acceptable salts or prodrugs thereof:

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a divisional of U.S. patent application Ser. No.12/240,678, filed on Sep. 29, 2008, which is a divisional of U.S. patentapplication Ser. No. 11/115,225, filed on Apr. 27, 2005, which claimsthe benefit of U.S. Provisional Application No. 60/565,527, filed Apr.27, 2004, the entire contents of which are incorporated herein byreference.

FIELD OF THE INVENTION

Method of treating interstitial cystitis irritable bowel syndrome,ulcerative colitis and other conditions involving inflammation.

BACKGROUND OF THE INVENTION

Interstitial cystitis (IC) is a chronic inflammation or irritation ofthe urinary bladder wall that is estimated to affect 700,000 people(approximately 90% female) in the United States. The primary symptoms ofIC are urinary urgency, frequency, and often-severe pelvic and perinealpain. The similarity of the symptoms of IC with other bladder diseases,such as urinary tract infections (UTI), urethritis, urethral syndrome,trigonitis, prostatitis, dysuria, and nocturia has caused difficulty inthe diagnosis of the disease. After other similar bladder diseases areruled out and a cystoscopic examination of the bladder wall revealscharacteristic signs of IC, including small petechial hemorrhages orlarger Hunner's Ulcers, IC is usually diagnosed.

The etiology and pathogenesis of the transmural inflammation in IC areunclear. The diagnosis is one of exclusion, and the treatment is usuallyempiric. The symptoms of IC arise from a defect in the glycosaminoglycan(GAG) component of the mucin layer that covers and protects the bladderurothelium. A deficiency of this layer is thought to cause IC.

The GAG hypothesis is the basis for treating interstitial cystitis withglycosaminoglycan “replacements,” such as sodium pentosan polysulfateand heparin or hyaluronic acid administered intravesically. Irritatingsubstances in the urine may leak through the urothelium, causinginflammation, tissue irritation and injury, mast cell degranulation andsensory nerve depolarization, which result in the urinary urgency,frequency and pain of interstitial cystitis. Glycosaminoglycans arehighly hydrophilic and protective. They maintain a stable layer of waterbetween the urothelium and the bladder lumen, and prevent adhesion toand invasion of the urothelium by bacteria, microcrystals, ions,proteins and other substances in the urine.

Currently there is no permanent cure for IC in the majority of patients.Treatment of IC with drug therapy has been proven to be the mosteffective means of alleviating symptoms. Oral medications for IC includebladder-coating agents, antidepressants, antihistamines, antispasmodics,and anesthetics. The effectiveness of oral medications is limited by thecirculating concentration of the drug in the blood stream. To addressthis limitation, many IC patients elect to undergo a procedure calledurinary bladder instillation, in which a therapeutic solution is pumpedinto the bladder through a urethral catheter.

Proposed treatments include pentosan polysulfate, anti-inflammatory orimmunosuppressant therapy, muscle relaxants, anti-histamines, andanalgesics. Of these, only pentosan polysulfate (Elmiron) has beenspecifically approved by the FDA for IC. None of the proposed therapies,including pentosan polysulfate, which may take up to 6 months to work,is universally accepted or universally efficacious. There is no cure andno effective treatment that works for everyone; treatments are intendedto relieve symptoms. Bladder distension, which is often part ofdiagnosis, is first line, because this treatment may amelioratesymptoms. Surgery is considered only in last resort and does not offersignificant long term benefit.

The related conditions of irritable bowel syndrome and ulcerativecolitis are also difficult to treat with existing therapeutic agents.These are nonspecific inflammatory diseases of the colon of unknowncause, characterized by diarrhea with discharge of mucus and blood,cramping abdominal pain, and inflammation and edema of the mucousmembrane with patches of ulceration.

Without meaning to be limited by theory, we believe that a uniquebreadth of receptor sites can be blocked by compounds of the presentinvention. Few, if any, of the known inhibitors of inflammatory diseaseembody all of the following sites of activity in a single molecule:inhibition of 1) leukotriene synthesis, 2) leukotriene D-4 receptors, 3)leukotriene E-4 receptors, 4) cAMP PDE III, 5) cAMP PDE IV, 6) synthesisof thromboxane A-2, 7) eosinophil migration and 8) lymphocyte migration.The above mechanisms are involved and cooperate in different degrees andwith different specificities among the wide variety of cells interactingin the so-called “inflammatory cascade,” to produce a fission-likeresult. By blocking a wide variety of action sites, compounds 1 and 2are expected to be effective for a wide array of illnesses that fallunder the umbrella of “inflammatory diseases.”

Since the biochemical messengers of inflammation cause an “explosive”type of reaction in the tissues, we suggest that the additive effects ofthe broad sites of action of the present compounds will result in high“multiplier” effects in amelioration of the urinary conditioninterstitial cystitis, as well as inflammatory disorders of thegastrointestinal tract, especially irritable bowel syndrome andulcerative colitis.

SUMMARY OF THE INVENTION

An object of the invention is to provide a method of treatinginterstitial cystitis, comprising: administering to a patient in need oftreatment for interstitial cystitis an effective amount of compound 1

or a pharmaceutically acceptable salt thereof.

Another object of the invention is to provide a method of treatinginterstitial cystitis comprising, administering an effective amount ofcompound 2, a metabolite of compound 1:

Another object of the invention is to provide a method of treatinginterstitial cystitis using an effective amount of compound 1 present inpolymorphic Form A.

Another object of the invention is to provide a method of treatingIBS/ulcerative colitis by administering an effective amount of compound1 or 2.

BRIEF DESCRIPTION OF THE FIGURES

FIGS. 1A-1E. Representative bladder CMGs during (A) saline infusion, (B)OA infusion, (C). OA infusion after Compound 1 (10 mg/kg); (D) OAinfusion after Compound 1 (30 mg/kg); (E) OA infusion after Compound 1(50 mg/kg).

FIGS. 2A-2D. Voiding frequency in (A) Control and OA sensitized rats;(B) before and after Compound 1 (10 mg/kg) treatment; (C) before andafter Compound 1 (30 mg/kg) treatment; (D) before and after Compound 1(50 mg/kg) treatment.

FIGS. 3A-3D. NVCs in (A) Control and OA sensitized rats; (B) before andafter Compound 1 (10 mg/kg) treatment; (C) before and after Compound 1(30 mg/kg) treatment; (D) before and after Compound 1 (50 mg/kg)treatment. An * denotes significance when compared to group 1. A ±denotes significance when compared to group 2.

FIG. 4. ICI in control and different treatment groups. An * denotessignificance when compared to group 1. A ± denotes significance whencompared to group 2.

FIG. 5. NVC in control and different treatment groups. An * denotessignificance when compared to group 1. A ± denotes significance whencompared to group 2.

DETAILED DESCRIPTION OF THE INVENTION

The compound4-[6-acetyl-3-[3-(4-acetyl-3-hydroxy-2-propylphenylthio)propoxy]-2-propylphenoxy]butyricacid (1) (MN-001), and its pharmaceutically acceptable salts andprotected prodrug forms, are orally active for treatment of interstitialcystitis.

The compound of formula 2 (MN-002), a metabolite of compound 1, is thedominant form in human plasma. It is also active for treatment ofinterstitial cystitis and can exist as two stereoisomers. The in vitropotency of the separated optical isomers of compound 2 are nearlyidentical against leukotrienes. Similarly, the acute toxicity (LD50) ofboth optical isomers is about the same. The isomers can be isolated bychiral chromatography methods. The racemate has been tested in long-termrodent and dog safety studies. Pharmacologically, the in vivo activityof the (−)-isomer and the racemate are similar. Pharmacokinetically, theisomers behave similarly in rats and dogs, with no evidence ofinterconversion.

The synthesis and biological activity of many phenoxyalkylcarboxylicacid derivatives, including compounds 1 and 2, are described by Ohashiet al. in U.S. Pat. Nos. 4,985,585 and 5,290,812, which are incorporatedherein by reference.Compound 1 may be prepared by reacting a phenol of formula 3:

wherein R is an acid protecting group, such as methyl or ethyl, with thebromo compound of formula 4 to give compound 5:

in an organic solvent, for example acetone, methylethylketone,diethylketone or dimethylformamide. The nucleophilic displacementreaction may be conducted from below room temperature up to the refluxtemperature of the solvent, in the presence of an inorganic base, e.g.,potassium carbonate or sodium carbonate. The addition of potassiumiodide is also recommended. Analogues of compound 4 having alternativeleaving groups, such as chloro and tosylate, may be used to effect thiscoupling reaction.

Removal of the protecting group in 5 by alkaline ester hydrolysis andextractive work-up gives compound 1 as a white solid.

Recrystallization of the white solid under controlled conditions givesan essentially pure polymorph, designated as Form A crystals (e.g., 90%or more, preferably at least 95% Form A). Polymorphic Form A andprocesses for producing it are described in U.S. patent application Ser.Nos. 10/601,861 and 10/601,862, both filed Jun. 24, 2003; which areincorporated herein by reference. All polymorphic forms of compound 1are active, but Form A is preferred. The solubility and thebioavailability of this polymorph, identified as orthorhombic crystals,is superior to the other polymorphs and thus Form A offers improvedsolid formulations.

Form A crystals can be obtained by dissolving compound 1 in 5 to 10parts by weight of ethanol at 25-40° C. to give a yellow to orangesolution. The ethanol solution is charged with 1-10 parts of water andagitated at 20-25° C. for about 15-60 minutes and then at 5-10° C. foran additional period of 1-4 hours, preferably 2.0-3.0 hours, resultingin an off-white suspension. To this suspension is added 5-15 parts ofwater and the mixture is agitated at 5-10° C. for an additional 1-4hours, preferably 1.5-2.0 hours. A solid, white to off-white product isisolated by vacuum filtration and the filter cake is washed with waterand dried in a vacuum at 25-40° C. for 12-24 hours.

Additional synthetic methods are shown below, where DHPA is2,4-dihydroxy-3-propylacetophenone and CHPA is4-(3-chloropropoxy)-2-hydroxyl-3-propylacetophenone.

Formulation and Administration

Administration of compounds 1 and 2, their mixtures and/orpharmaceutically acceptable salts or prodrugs can be done orally, or byintravenous, intramuscular, and subcutaneous injection, or transdermalmethods. Effective dosage levels can vary widely, e.g., from about 100to 2500 mg per day. The preferred daily dosage range is 250 to 2,000 mg,give in one, two or three portions. A preferred dose for IC is 1000 mgtwice a day. Suitable doses for treatment of IBS/ulcerative colitisinclude 1000 mg qd, 1000 mg bid, and 750 mg tid. Actual amounts willdepend on the circumstances of the patient being treated. As thoseskilled in the art recognize, many factors that modify the action of theactive substance will be taken into account by the treating physiciansuch as the age, body weight, sex, diet and condition of the patient,the time of administration, the rate and route of administration.Optimal dosages for a given set of conditions can be ascertained bythose skilled in the art using conventional dosage determination tests.

Suitable salts are sodium, potassium, calcium and magnesium, with sodiumbeing preferred. Suitable prodrug forms include carboxylic esters whichare readily hydrolyzable in vivo and release the active acid in vivo. Anexample of such a protecting group is disclosed in U.S. Pat. No.4,428,935, incorporated herein by reference.

The following protecting group is described in U.S. Pat. No. 4,428,935:

whereinR is hydrogen;Q is selected from the group consisting ofN(R¹)(R²)(R³) wherein R¹, R² and R³ are each alkyl having 1-4 carbonatoms, hydroxyalkyl having 1-4 carbon atoms, alkoxycarbonylalkyl whereinthe alkyl has 1-4 carbon atoms and the alkoxy has 1-4 carbon atoms oraminocarbonylalkyl wherein the alkyl has 1-4 carbon atoms,N-alkyl-morpholyl wherein the alkyl has 1-4 carbon atoms,N-alkyl-piperidyl wherein the alkyl has 1-4 carbon atoms orN-alkyl-pyrrolidyl wherein the alkyl has 1-4 carbon atom, and thosegroups substituted at a ring position with alkyl having 1-4 carbonatoms, aminocarbonyl, hydroxyl or alkoxycarbonyl wherein the alkoxy has1-4 carbon atoms,bicyclo [2.2.2]1-,4-diazaoctanyl and those substituted with alkyl having1-4 carbon atoms, aminocarbonyl, hydroxyl or alkoxycarbonyl wherein thealkoxy has 1-4 carbon atoms,pyridyl, alkylpyridyl wherein the alkyl has 1-4 carbon atoms,dialkylpyridyl wherein each alkyl has 1-4 carbon atoms, trialkylpyridylwherein each alkyl has 1-4 carbon atoms,halopyridyl, cyanopyridyl, nitropyridyl, aminocarbonylpyridyl,N-alkylaminocarbonylpyridyl wherein the alkyl has 1-4 carbon atoms,N,N-dialkylaminocarbonylpyridyl wherein each alkyl has 1-4 carbon atoms,alkoxycarbonylpyridyl wherein the alkoxy contains 1-4 carbon atoms, orN,N-dialkylaminopyridyl wherein each alkyl has 1-4 carbon atoms, andquinolyl or N-alkylimidazolyl wherein the alkyl has 1-4 carbon atoms;andX⁻ is a pharmaceutically-acceptable anion.

The compounds of the present invention can be formulated in anypharmaceutically acceptable form, including liquids, powders, creams,emulsions, pills, troches, suppositories, suspensions, solutions, andthe like. Therapeutic compositions containing compounds 1 and 2, theirmixtures and/or pharmaceutically acceptable salts will ordinarily beformulated with one or more pharmaceutically acceptable ingredients inaccordance with known and established practice. In general, tablets areformed utilizing a carrier such as modified starch, alone or incombination with 10% by weight of carboxymethyl cellulose (Avicel). Theformulations are compressed at from 1,000 to 3,000 pounds pressure inthe tablet-forming process. The tablets preferably exhibit an averagehardness of about 1.5 to 8.0 kp/cm², preferably 5.0 to 7.5 kp/cm².Disintegration time varies from about 30 seconds to about 15 or 20minutes. The following examples give specific embodiments of theinvention but should not be construed as limiting its scope.

Formulations for oral use can be provided as hard gelatin capsuleswherein the phenoxyalkylcarboxylic acid, mixtures thereof orpharmaceutically acceptable salts are mixed with an inert solid diluentsuch as calcium carbonate, calcium phosphate or kaolin, or as softgelatin capsules in which the compounds are mixed with an oleaginousmedium, e.g., liquid paraffin or olive oil. Suitable carriers includemagnesium carbonate, magnesium stearate, talc, sugar, lactose, pectin,dextrin, starch, gelatin, tragacanth, methylcellulose, sodiumcarboxymethylcellulose, a low melting wax, cocoa butter, and the like.

Aqueous suspensions can contain the compounds, their mixtures and/orpharmaceutically acceptable salts in admixture with pharmaceuticallyacceptable excipients such as suspending agents, e.g., sodiumcarboxymethyl cellulose, methylcellulose, hydroxypropylmethylcellulose,sodium alginate, polyvinylpyrrolidone, gum tragacanth and gum acacia;dispersing or wetting agents such as naturally occurring phosphatide,e.g., lecithin, or condensation products of an alkaline oxide with fattyacids, e.g., polyoxyethylene stearate, or condensation products ofethylene oxide with long chain aliphatic alcohols, e.g,heptadecaethylene-oxycetanol, or condensation products of ethylene oxidewith partial esters derived from fatty acids and a hexitol, e.g.,polyoxyethylene sorbitol monoleate or condensation products of ethyleneoxide with partial esters derived from fatty acids and hexitolanhydrides, e.g., polyoxyethylene sorbitan monoleate. Such aqueoussuspensions can also contain one or more preservatives, e.g.,ethyl-or-n-propyl-p-hydroxy benzoate, one or more coloring agents, oneor more flavoring agents and one or more sweetening agents, such assucrose, saccharin or sodium or calcium cyclamate.

Dispersible powders and granules suitable for preparation of an aqueoussuspension by the addition of water provide the compounds of formulas 1and 2, their mixtures and/or pharmaceutically acceptable salts inadmixture with a dispersing or wetting agent, suspending agent and oneor more preservatives. Suitable dispersing or wetting agents andsuspending agents are exemplified by those already mentioned above.Additional excpients, e.g., sweetening, flavoring and coloring agents,can also be present. Syrups and elixirs can be formulated withsweetening agents, for example glycerol, sorbitol or sucrose. Suchformulations can also contain a demulcent, a preservative and flavoringand coloring agents.

The present invention can be administered in sustained release dosageform, of which many kinds are known, e.g., as described in U.S. Pat.Nos. 4,788,055; 4,816,264; 4,828,836; 4,834,965; 4,834,985; 4,996,047;5,071,646; and, 5,133,974, the contents of which are incorporated byreference herein.

It is also within the scope of this invention to administer the subjectcompounds concurrently with, or after administration of any other knownpharmacologically active agent useful for treating urinary incontinence.Such agents include, anticholinergics such as oxybutynin, atropine,propantheline, terodiline, and dicyclomine; sympathomimetics such asephedrine, pseudoephedrine, epinephrine, and phenylpropanolamine;tricyclic antidepressants such as imipramine, doxepin, andamitriptyline; estrogens or estrogen-related compounds havingestrogen-like activity such as estradiol, and estrone; andantispasmodics or direct acting bladder smooth muscle relaxants such asflavoxate. For a detailed discussion of these pharmacologically activeagents, reference may be made to “Goodman and Gillman's PharmacologicalBasis of Therapeutics”, Goodman et al., eds. 7th ed., 1985, Macmillanand Company, New York.

Other forms suitable for oral administration include liquid formpreparations including emulsions, syrups, elixirs, aqueous solutions,aqueous suspensions, or solid form preparations which are intended to beconverted shortly before use to liquid form preparations. Emulsions maybe prepared in solutions, for example, in aqueous propylene glycolsolutions or may contain emulsifying agents, for example, such aslecithin, sorbitan monooleate, or acacia. Aqueous solutions can beprepared by dissolving the active component in water and adding suitablecolorants, flavors, stabilizing, and thickening agents. Aqueoussuspensions can be prepared by dispersing the finely divided activecomponent in water with viscous material, such as natural or syntheticgums, resins, methylcellulose, sodium carboxymethylcellulose, and otherwell known suspending agents. Solid form preparations include solutions,suspensions, and emulsions, and may contain, in addition to the activecomponent, colorants, flavors, stabilizers, buffers, artificial andnatural sweeteners, dispersants, thickeners, solubilizing agents, andthe like.

The compounds of the present invention may be formulated for parenteraladministration (e.g., by injection, for example bolus injection orcontinuous infusion) and may be presented in unit dose form in ampoules,pre-filled syringes, small volume infusion or in multi-dose containerswith an added preservative. The compositions may take such forms assuspensions, solutions, or emulsions in oily or aqueous vehicles, forexample solutions in aqueous polyethylene glycol. Examples of oily ornonaqueous carriers, diluents, solvents or vehicles include propyleneglycol, polyethylene glycol, vegetable oils (e.g., olive oil), andinjectable organic esters (e.g., ethyl oleate), and may containformulatory agents such as preserving, wetting, emulsifying orsuspending, stabilizing and/or dispersing agents. Alternatively, theactive ingredient may be in powder form, obtained by aseptic isolationof sterile solid or by lyophilisation from solution for constitutionbefore use with a suitable vehicle, e.g., sterile, pyrogen-free water.

The compounds of the present invention may be formulated for topicaladministration to the epidermis as ointments, creams or lotions, or as atransdermal patch. Ointments and creams may, for example, be formulatedwith an aqueous or oily base with the addition of suitable thickeningand/or gelling agents. Lotions may be formulated with an aqueous or oilybase and will in general also containing one or more emulsifying agents,stabilizing agents, dispersing agents, suspending agents, thickeningagents, or coloring agents. Formulations suitable for topicaladministration in the mouth include lozenges comprising active agents ina flavored base, usually sucrose and acacia or tragacanth; pastillescomprising the active ingredient in an inert base such as gelatin andglycerin or sucrose and acacia; and mouthwashes comprising the activeingredient in a suitable liquid carrier.

The compounds of the present invention may be formulated foradministration as suppositories. A low melting wax, such as a mixture offatty acid glycerides or cocoa butter is first melted and the activecomponent is dispersed homogeneously, for example, by stirring. Themolten homogeneous mixture is then poured into convenient sized molds,allowed to cool, and to solidify.

The compounds of the present invention may be formulated for vaginaladministration. Pessaries, tampons, creams, gels, pastes, foams orsprays containing in addition to the active ingredient such carriers asare known in the art to be appropriate.

The compounds of the present invention may be formulated for nasaladministration. The solutions or suspensions are applied directly to thenasal cavity by conventional means, for example, with a dropper, pipetteor spray. The formulations may be provided in a single or multidoseform. In the latter case of a dropper or pipette, this may be achievedby the patient administering an appropriate, predetermined volume of thesolution or suspension. In the case of a spray, this may be achieved forexample by means of a metering atomizing spray pump.

The compounds of the present invention may be formulated for aerosoladministration, particularly to the respiratory tract and includingintranasal administration. The compound will generally have a smallparticle size for example of the order of five (5) microns or less. Sucha particle size may be obtained by means known in the art, for exampleby micronization. The active ingredient is provided in a pressurizedpack with a suitable propellant such as a chlorofluorocarbon (CFC), (forexample, dichlorodifluoromethane, trichlorofluoromethane, ordichlorotetrafluoroethane), carbon dioxide or other suitable gas. Theaerosol may conveniently also contain a surfactant such as lecithin. Thedose of drug may be controlled by a metered valve. Alternatively theactive ingredients may be provided in a form of a dry powder, forexample a powder mix of the compound in a suitable powder base such aslactose, starch, starch derivatives such as hydroxypropylmethylcellulose and polyvinylpyrrolidine (PVP). The powder carrier will form agel in the nasal cavity. The powder composition may be presented in unitdose form for example in capsules or cartridges of e.g., gelatin orblister packs from which the powder may be administered by means of aninhaler.

When desired, formulations can be prepared with enteric coatings adaptedfor sustained or controlled release administration of the activeingredient. A common type of controlled-release formulation that may beused for the purposes of the present invention comprises an inert core,such as a sugar sphere, coated with an inner drug-containing layer andan outer membrane layer controlling drug release from the inner layer.

The cores are preferably of a water-soluble or swellable material, andmay be any such material that is conventionally used as cores or anyother pharmaceutically acceptable water-soluble or water-swellablematerial made into beads or pellets. The cores may be spheres ofmaterials such as sucrose/starch (Sugar Spheres NF), sucrose crystals,or extruded and dried spheres typically comprised of excipients such asmicrocrystalline cellulose and lactose.

The substantially water-insoluble material in the first layer isgenerally a “GI insoluble” or “GI partially insoluble” film formingpolymer (dispersed or dissolved in a solvent). As examples may bementioned ethyl cellulose, cellulose acetate, cellulose acetatebutyrate, polymethacrylates such as ethyl acrylate/methyl methacrylatecopolymer (Eudragit NE-30-D) and ammonio methacrylate copolymer types Aand B (Eudragit RL30D and RS30D), and silicone elastomers. Usually, aplasticizer is used together with the polymer. Exemplary plasticizersinclude: dibutylsebacate, propylene glycol, triethylcitrate,tributylcitrate, castor oil, acetylated monoglycerides, acetyltriethylcitrate, acetyl butylcitrate, diethyl phthalate, dibutylphthalate, triacetin, fractionated coconut oil (medium-chaintriglycerides).

The second layer containing the active ingredient may be comprised ofthe active ingredient (drug) with or without a polymer as a binder. Thebinder, when used, is usually hydrophilic but may be water-soluble orwater-insoluble. Exemplary polymers to be used in the second layercontaining the active drug are hydrophilic polymers such aspolyvinylpyrrolidone (PVP), polyalkylene glycol such as polyethyleneglycol, gelatine, polyvinyl alcohol, starch and derivatives thereof,cellulose derivatives, such as hydroxypropylmethyl cellulose (HPMC),hydroxypropyl cellulose, carboxymethyl cellulose, methyl cellulose,ethyl cellulose, hydroxyethyl cellulose, carboxyethyl cellulose,carboxymethylhydroxyethyl cellulose, acrylic acid polymers,polymethacrylates, or any other pharmaceutically acceptable polymer. Theratio of drug to hydrophilic polymer in the second layer is usually inthe range of from 1:100 to 100:1 (w/w).

Suitable polymers for use in the third layer, or membrane, forcontrolling the drug release may be selected from water-insolublepolymers or polymers with pH-dependent solubility, such as, for example,ethyl cellulose, hydroxypropylmethyl cellulose phthalate, celluloseacetate phthalate, cellulose acetate trimellitate, polymethacrylates, ormixtures thereof, optionally combined with plasticizers, such as thosementioned above.

Optionally, the controlled release layer comprises, in addition to thepolymers above, another substance(s) with different solubilitycharacteristics, to adjust the permeability, and thereby the releaserate, of the controlled release layer. Exemplary polymers that may beused as a modifier together with, for example, ethyl cellulose include:HPMC, hydroxyethyl cellulose, hydroxypropyl cellulose, methylcellulose,carboxymethylcellulose, polyethylene glycol, polyvinylpyrrolidone (PVP),polyvinyl alcohol, polymers with pH-dependent solubility, such ascellulose acetate phthalate or ammonio methacrylate copolymer andmethacrylic acid copolymer, or mixtures thereof. Additives such assucrose, lactose and pharmaceutical grade surfactants may also beincluded in the controlled release layer, if desired.

The pharmaceutical preparations are preferably in unit dosage forms. Insuch form, the preparation is subdivided into unit doses containingappropriate quantities of the active component. The unit dosage form canbe a packaged preparation, the package containing discrete quantities ofpreparation, such as packeted tablets, capsules, and powders in vials orampoules. Also, the unit dosage form can be a capsule, tablet, cachet,or lozenge itself, or it can be the appropriate number of any of thesein packaged form.

Other suitable pharmaceutical carriers and their formulations aredescribed in Remington: The Science and Practice of Pharmacy 1995,edited by E. W. Martin, Mack Publishing Company, 19th edition, Easton,Pa. Representative pharmaceutical formulations containing a compound ofthe present invention are described in the Examples.

The following table reports data from a comprehensive biochemicalevaluation of compounds 1 and 2, showing these drugs can hit severalinflammatory chemical mediators simultaneously, which appears to be thebasis for their unique therapeutic activities. TNF alpha has beensuggested as a probable disease provocateur in arthritis, heart disease,IBD and cachexia. Likewise, the cAMP/cGMP system is implicated in a widenumber of diseases or disorders and are expected to respond to treatmentwith compounds 1 and 2 including prostatitis, IBS/IBD, and multiplesclerosis, etc.

IC₅₀ (K_(i)) IC₅₀ (K_(i)) (μM) (μM) Biochemical Assay Species MN-001MN-002 LTC4 Synthetase Gp 24.7 18.3 5-LO Hu 1.11 1.61 PDE1 Bov 0.7413.08 PDE2 Hu 3.43 3.38 PDE3 Hu 0.567 1.68 PDE4 Hu 0.639 0.303 PDE5 Hu12.7 8.47 PDE6 Bov 2.05 1.73 PLA2-I Pig 142 248 PLC Ba 0.59 5.13Adrenergic α2B Hu 2.81 (1.28)  1.6 (0.732) Adrenergic β3 Hu 4.63 (3.47) 1.1 (0.821) Adrenergic, NE Transporter Hu  3.5 (3.47) 2.05 (2.03) LTB4Hu 9.02 (2.29) — LTD4 Gp 6.36 (3.18) 5.35 (2.67) Melanocortin MC5 Hu2.58 (2.42) 1.82 (1.71) Opiate δ Hu  1.04 (0.365) 3.03 (1.07)Thromboxane A2 Hu 3.37 (2.2)  3.89 (2.54) Ba = bacillus cereus, — = nottested

The following examples are illustrative of the present invention andshould not be construed as limiting.

Example 1 Synthesis of ethyl4-[6-acetyl-3-[3-(4-acetyl-3-hydroxy-2-propylphenylthio)-propoxy]-2-propylphenoxy]butyrate

To a stirred mixture of ethyl4-(6-acetyl-3-hydroxy-2-propylphenoxy)butyrate (1.6 g), potassium iodide(0.5 g) and potassium carbonate (1.45 g) in acetone (30 ml) was addeddrop wise a solution of4-(3-bromopropylthio)-2-hydroxy-3-propylphenyl-ethanone (1.9 g) inacetone (10 ml) with heating to reflux. After refluxing six hours themixture was cooled to room temperature and inorganic materials wereseparated by filtration. The filtrate was concentrated and the residuewas separated and purified by silica-gel column chromatography (elutingwith benzene:ethyl acetate=9:1) to give the title compound as crudecrystals (2.1 g, 72.4%) which were recrystallized from ethanol to givecolorless crystals, mp 65-66° C.

Example 2 Synthesis of4-[6-acetyl-3-[3-(4-acetyl-3-hydroxy-2-propylphenylthio)propoxy]-2-propylphenoxy]butyricacid

To a mixture of ethyl4-[6-acetyl-3-[3-(4-acetyl-3-hydroxy-2-propylphenylthio)propoxy]-2-propylphenoxy]butyrate(2.1 g) in ethanol (10 ml) was added a solution of sodium hydroxide(0.26 g) dissolved into water (10 ml). After heating on a hot water bathfor 5 minutes, the mixture was cooled by adding ice-water and was madeacidic by addition of hydrochloric acid, followed by extraction withethyl acetate. The organic layer was washed with water, dried oversodium sulfate and concentrated. The resultant residue was separated andpurified by silica-gel column chromatography (eluting withethanol:methylene chloride=3:100) to give the title compound (1.3 g,65.2%) as colorless crystals, mp 79-81° C.

Example 3

Bulk crystallization procedure for obtaining orthorhombic polymorph,crystal type V (Form A).

Off-white solid compound (1) 34 g was dissolved in 204 mL (6 parts wrtmass of dry filter cake) of ethanol (40° C.) giving a yellow to orangesolution. With moderate agitation, the ethanol solution was charged with43 mL (1.3 parts) of water. The reaction mixture was cooled to 20-25° C.and agitated at 20-25° C. for about 15 minutes and then at 10-15° C. foran additional period of 1-2 hours, appearing as an off-white suspension.

To the resulting suspension was then charged 364 mL (10.7 parts) ofwater and the mixture was agitated at 5-10° C. for an additional 1-2hours. A solid, white to off-white product was isolated by vacuumfiltration. The filter cake was washed with 2×30 mL of water. The offwhite solid was dried in a vacuum at 35-40° C. for 24 hours.

Example 4

In general, wet granulation tablets were prepared with a bindingsolution comprised of an aqueous solution of hydroxypropylcellulose.Granulation was performed with a high shear granulator, the resultantwet mass was fluid bed dried, milled, blended with extragranularexcipients to aid disintegration, flow and compressibility, andsubsequently tabletted on a tablet press. These core tablets were filmcoated to standardize appearance and to improve compliance (i.e. ease ofswallowing). Excipients included, but were not limited to croscarmellosesodium, magnesium stearate, hydroxypropylcelluse,hydroxypropylmethylcellulose, lactose, glyceryl behenate,polyvinylpyrrolidine, mannitol, titanium dioxide and microcrystallinecellulose.

Example 5

In general, the dry granulation formulation was formed by dry blending(in a tumble blender or high shear mixer) a portion of the binding,disintegration and lubrication powders. This dry powder blend was formedinto granules through the use of a roller compactor equipped with anoscillating (shear) granulator. The ss mesh screen, gap width, gapforce, roller speed and granulator speeds were defined to optimize theformulation physical parameters as apparent to those skilled in the artof pharmaceutical processing. Excipients included, but were not limitedto croscarmellose sodium, magnesium stearate, hydroxypropylcelluse,hydroxypropylmethylcellulose, lactose, glyceryl behenate,polyvinylpyrrolidine, mannitol, titanium dioxide and microcrystallinecellulose.

Example 6 Specific Formulation for Dry Granulation

Prototype 1 Prototype 2 No. Ingredient (mg/tablet) (mg/tablet) 1Compound (1) (Form A) 250 250 2 Lactose regular/fast flow 7.5 — 3Microcrystalline cellulose PH101 31 31 4 Croscarmellose sodium 20 20 5Hydroxypropylcellulose 80 — 6 Magnesium stearate 2.0 — 7Hydroxypropylmethylcellulose 2910 8.0 — 8 Titanium Dioxide 1.0 — 9Carnauba wax 0.5 0.5 10 Polyvinylpyrrolidone — 85 11 Mannitol — 3.5 12Glyceryl behenate — 2.0  13 Opadry II (white) — 8.0 Total 400 mg 400 mg

Example 7 Specific Formulations for Wet Granulation

Prototype 3 Prototype 4 No. Ingredient (mg/tablet) (mg/tablet) 1Compound (1) (Form A) 250 250 2 Lactose regular/fast flow 7.5 — 3Microcrystalline cellulose PH101 32 32 4 Croscarmellose sodium 25 25 5Hydroxypropylcellulose 25 — 6 Magnesium stearate 2.0 — 7Hydroxypropylmethylcellulose 2910 7.0 — 8 Titanium Dioxide 1.0 — 9Carnauba wax 0.5 0.5 10 Polyvinylpyrrolidone — 30 11 Mannitol — 3.5 12Glyceryl behenate — 2.0 13 Opadry II (white) — 7.0 Total 350 mg 350 mg

Example 8

A 35-year-old female complains of suffering symptoms associated withinterstitial cystitis (IC), including gastrointestinal symptoms ofdiarrhea, bloating and pain. In an effort to alleviate these symptoms, atablet containing 1000 mg of compound (1) (Form A) is administeredorally to this female patient twice daily for at least 2 months. Afterthis time period, this patient reports positive effects, including areduction in the symptoms she complained of. Similar positive resultsare observed for other patients complaining of the same or similarsymptoms attributable to IC.

Example 9

A 35-year-old male complains of symptoms associated with IBS/ulcerativecolitis, including diarrhea with discharge of mucus and blood, crampingabdominal pain, and inflammation and edema of the mucous membrane withpatches of ulceration. The patient is treated with tablets containing750 mg of compound (1), three times per day, for at least two months.After this time period, he reports positive effects, including areduction in the symptoms complained of. Similar positive results areobserved for other patients complaining of the same or similar symptomsattributable to IBS or ulcerative colitis.

Example 10

Previous reports have demonstrated that intravesical instillation ofovalbumin in sensitized guinea pigs increases bladder contractions. See,Ahluwalia A, Giuliani S, Scotland R and Maggi C A: Ovalbumin-inducedneurogenic inflammation in the bladder of sensitized rats. Br. J.Pharmacol. (1998) 124:190-6. Compound 1 (1), exhibitinganti-inflammatory activity with mast cell stabilizing potential, hasbeen shown to be beneficial in suppressing ovalbumin-induced bladdercontractions. The protective potential of 1 in a rat model ofinflammation-induced bladder hyperactivity has been evaluated, asdescribed below. This rat model serves as an animal model forinterstitial cystitis, irritable bowel syndrome, ulcerative colitis, andpotentially other inflammatory conditions.

Methods: A. Sensitization of Rats:

Sprague-Dawley rats (n=50; 200±250 g) were utilized for this study. Theanimals were divided into 5 groups (n=10). The first group served as thecontrol and the second group was sensitized with ovalbumin (OA). Thethird, fourth, and fifth groups were also sensitized as described butsubjected to oral gavaging with Compound 1 prior to acute ovalbuminchallenge.

Sensitization of the animals was accomplished with an intraperitonealinjection of a mixture of 1 mg OA and 100 mg aluminum hydroxidesuspended in 1 mL of saline. Fourteen days later, these sensitized ratswere anaesthetized with a subcutaneous injection of urethane (1.2 g/kg)for intravesical OA (10 mg/mL) administration and evaluation of bladderhyperactivity. The animals in group 1 received saline (control), group 2animals received about 2 mL of OA (10 mg/mL OA in sterile saline) andanimal groups 3-5 animals received oral 1 (10, 30, 50 mg/kg in gumacacia suspension) 60 minutes prior to acute OA challenge (2 mL of 10mg/mL OA).

B. Evaluation of Bladder Overactivity:

All animals were subjected to evaluation of bladder hyperactivity.Briefly, a 1 cm incision was made along the centerline of the lowerventral abdomen. The bladder was exteriorized, and catheterized by meansof a polyethylene tube (PE 50, Clay Adams) inserted into the bladderdome and sutured in place using a 2-0 braided silk suture. The bladderwas returned to the abdomen, with the catheter line escaping through theincision. The catheter was then connected to a pressure transducer (UFI,Morro Bay, Calif.) and in turn, connected to an infusion pump (HarvardApparatus, MA). During the continuous filling bladder cystometry, thepressure was recorded with the transducer using the program LabVIEW(National Instruments, TX).

For cystometry (See, Chuang Y C, Chancellor M B, Seki S, Yoshimura N,Tyagi P, Huang L, Lavelle J P, De Groat W C and Fraser M O: Intravesicalprotamine sulfate and potassium chloride as a model for bladderhyperactivity. Urology (2003) 61:664-70), the bladder was first infusedwith warm 0.9% saline (37° C.) at 40 μL/min (2.4 mL/hr) and at least 20minutes of stable voiding cycles were recorded during infusion. Thisprocess was followed by intravesical infusion of OA (10 mg/mL) andbladder contractions were recorded. Frequency of contractions (voids),inter-contractile interval (ICI), and non-voiding contractions (NVC)were calculated from these recordings.

Results:

Representative tracings of bladder cystometrograms (CMGs) of treatmentgroups are depicted in FIG. 1A-1E. Infusion of intact rat bladder with0.9% saline resulted in normal and comparable numbers of voids and NVC(FIG. 1; Table I). In OA sensitized rats, intravesical OA (10 mg/mL)infusion resulted in a significant increase in NVC relative tonon-sensitized saline infusion values (FIGS. 1, 3 and 5; Table I).Pre-treatment with Compound 1 resulted in dose-dependent inhibition ofOA-induced changes in NVC and ICI (FIGS. 3, 4 and 5; Table I). Nosignificant differences were observed in voiding frequency (FIG. 2)between control (group 1) and different treatment groups (2-5).

CONCLUSIONS

These studies clearly indicate that acute intravesical challenge ofOA-sensitized rats causes contractions of bladder smooth muscle leadingto a significant increase in NVC and a decrease in ICI. Pre-treatmentwith 1, preferably at 30 and 50 mg/kg dose levels, produced asignificant protection against these OA-induced changes. Similaroutcomes are expected with the pre-treatment of Compound 2, a metaboliteof 1.

TABLE I Effect of intravesical ovalbumin on rat bladder before and aftertreatment with Compound 1 Statistical Agent infused into rat bladderNVC/minute significance* Group 1: Sodium Chloride (baseline) 0.2400 ±0.07483 — (n = 10) Group 2: OA (10 mg/mL) 0.9400 ± 0.2473 P < 0.05 (n =10) compared to Group 1 Group 3: OA (10 mg/mL) after giving 1 0.4700 ±0.1221 NS (10 mg/kg) compared (n = 10) to Group 2 Group 4: OA (10 mg/mL)after giving 1 0.3444 ± 0.06894 P < 0.05 (30 mg/kg) compared (n = 9) toGroup 2 Group 5: OA (10 mg/mL) after giving 1 0.1778 ± 0.08296 P < 0.05(50 mg/kg) compared (n = 9) to Group 2 *NS = not significant.

All documents cited herein are incorporated by reference in theirentirety. While the invention has been described in detail, and withreference to specific embodiments, it will be apparent to one withordinary skill in the art that various changes and modifications can bemade therein without departing from the spirit and scope of theinvention.

1. A method of treating irritable bowel syndrome in a patient sufferingtherefrom, comprising: administering an effective amount of compound (1)

or a pharmaceutically acceptable salt or an ester thereof.
 2. The methodof claim 1, wherein the compound is administered orally.
 3. The methodof claim 2, wherein the dosage form is a tablet or capsule.
 4. Themethod of claim 1, wherein the compound is present in polymorphic Form Aand is substantially free of other polymorphic forms.
 5. The method ofclaim 1, wherein the compound is administered in a liquid.
 6. The methodof claim 1, wherein the dose is from 250 to 2,500 mg/day, divided intoone, two or three portions.
 7. The method of claim 1, wherein thecompound is present as orthorhombic crystals.
 8. The method of claim 1,further comprising administration of compound (2)

or a pharmaceutically acceptable salt or an ester thereof.
 9. A methodof treating irritable bowel syndrome in a patient suffering therefrom,comprising: administering an effective amount of compound (2)

or a pharmaceutically acceptable salt or an ester thereof.